User:Boris Brumshtein

Personal Information
Name: Boris Brumshtein

Languages: English, Hebrew and Russian

Current residence: Rehovot, Israel.

Contact Information
Department of Structural Biology, Weizmann Institute of Science Rehovot, 76100, Israel

E-mail:	 Boris.Brumshtein@weizmann.ac.il 	Tell:  	+972-52-6902631 (private) +972-8-9342647 (work)

Education
Estimated graduation - summer 2008. Scientific advisors: Prof. Joel L. Sussman, Prof. Israel Silman, Prof. Anthony H. Futerman Thesis topic: "Structural and biochemical studies on acid-beta-glucosidase, the defective protein in Gaucher disease". Scientific advisor: Prof. Joel Sussman.
 * 2003 - present Direct PhD program at the Weizmann Institute of Science, Faculty of Chemistry, Department of Structural Biology.
 * 2002-2003	MSc student, Weizmann Institute of Science, Faculty of Chemistry, Department of Structural Biology.
 * 1999-2002	BA in Biochemistry, Technion, Israel Institute of Science and Technology, Faculty of Chemistry, cum laude.

Qualifications
In my research I employ different protein biochemistry, enzymology and crystallography methods, thus I gained a lot of experience in the following techniques:

Crystallography

 * Protein crystallization utilizing microbatch crystallization, IMPAX I-5, Oryx and Mosquito crystallization robots.
 * Ab initio crystallization and crystals optimization. Various crystallization techniques, such as: soaking, co-crystallization, crystallization in presence of heavy metals for phasing, etc.
 * Crystal handling and manipulations, micro-manipulations with ZEISS microscope/manipulator system.
 * Data acquisition using Rigaku Raxis 4/4++ or synchrotron x-ray sources.
 * X-ray data processing software: XDS, HKL2000.
 * Protein structure determination using MR, SIR, MIR, MAD, SIRAS approaches with CNS, SHELX and CCP4 software suits.
 * Molecular modeling, visualization and analysis software: PyMol, Coot, XtalView, O, CCP4, APBS.
 * Docking and small molecule/drug database mining with AutoDock - drug docking and virtual screening, Cambridge Structure Database (CSB), ZINC, ChemOffice.

Molecular biology and biochemistry

 * Protein cloning and expression in E.Coli.
 * Biochemical and protein purification techniques: HPLC, FPLC, Ni-columns, Ion exchange columns, acryl-amide gels, size exclusion, affinity chromatography, gradient and analytical ultracentrifugation, ultra-filtration, Mini Prep cell, etc.
 * Drug-Protein interaction analysis employing various structural, biochemical and biophysical assays such as: enzymatic assays, ELIZA, circular dichroism, dynamic light scattering, mass-spec, BiaCore, analytical ultracentrifugation, etc.

Programming languages

 * Unix, C, Perl, Matlab, Java (basic programming).

LIST OF PUBLICATIONS

 * 1) Kacher Y, Brumshtein B, Boldin-Adamsky S, Toker L, Shainskaya A, Silman I, Sussman JL, Futerman AH. Acid-beta-glucosidase: insights from structural analysis and relevance to Gaucher disease therapy. Biol Chem. 2008 Sep 11. PMID:18783340
 * 2) Harel M, Brumshtein B, Meged R, Dvir H, Ravelli RB, McCarthy A, Toker L, Silman I, Sussman JL. 3-D structure of serum paraoxonase 1 sheds light on its activity, stability, solubility and crystallizability. Arh Hig Rada Toksikol. 2007 Sep;58(3):347-53. PMID:17913690
 * 3) Brumshtein B, Greenblatt HM, Butters TD, Shaaltiel Y, Aviezer D, Silman I, Futerman AH, Sussman JL. Crystal structures of complexes of N-butyl- and N-nonyl-deoxynojirimycin bound to acid-beta -glucosidase: Insights into the mechanism of chemical chaperone action in Gaucher disease. J.Biol. Chem., Vol. 282, Issue 39, 29052-29058, (2007). PMID:17666401
 * 4) Brumshtein, B., Wormald, M. R., Silman, I., Futerman, A. H. & Sussman, J. L. Structural comparison of differently glycosylated forms of acid-beta-glucosidase, the defective enzyme in Gaucher disease. Acta Crystallogr D Biol Crystallogr 62, 1458-65 (2006). PMID:17139081
 * 5) Rydberg EH, Brumshtein B, Greenblatt HM, Wong DM, Shaya D, Williams LD, Carlier PR, Pang YP, Silman I, Sussman JL. Complexes of alkylene-linked tacrine dimers with Torpedo californica acetylcholinesterase: Binding of Bis5- tacrine produces a dramatic rearrangement in the active-site gorge. J Med Chem 49, 5491-500 (2006). PMID:16942022
 * 6) Harel M, Hyatt JL, Brumshtein B, Morton CL, Wadkins RM, Silman I, Sussman JL, Potter PM. The 3D structure of the anticancer prodrug CPT-11 with Torpedo californica acetylcholinesterase rationalizes its inhibitory action on AChE and its hydrolysis by butyrylcholinesterase and carboxylesterase. Chem Biol Interact 157-158, 153-7 (2005). PMID:16289500
 * 7) Hyatt JL, Tsurkan L, Morton CL, Yoon KJ, Harel M, Brumshtein B, Silman I, Sussman JL, Wadkins RM, Potter PM. Inhibition of acetylcholinesterase by the anticancer prodrug CPT-11. Chem Biol Interact 157-158, 247-52 (2005). PMID:16257398
 * 8) Harel M, Hyatt JL, Brumshtein B, Morton CL, Yoon KJ, Wadkins RM, Silman I, Sussman JL, Potter PM. The crystal structure of the complex of the anticancer prodrug 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin (CPT- 11) with Torpedo californica acetylcholinesterase provides a molecular explanation for its cholinergic action. Mol Pharmacol 67, 1874-81 (2005).  PMID:15772291
 * 9) Harel M, Aharoni A, Gaidukov L, Brumshtein B, Khersonsky O, Meged R, Dvir H, Ravelli RB, McCarthy A, Toker L, Silman I, Sussman JL, Tawfik DS. Structure and evolution of the serum paraoxonase family of detoxifying and anti-atherosclerotic enzymes. Nat Struct Mol Biol 11, 412-9 (2004). PMID:15098021
 * 10) Adir, N., Rukhman, V., Brumshtein, B. & Anati, R. Preliminary X-ray crystallographic analysis of a soluble form of MntC, a periplasmic manganese- binding component of an ABC-type Mn transporter from Synechocystis sp. PCC 6803. Acta Crystallogr D Biol Crystallogr 58, 1476-8 (2002). PMID:12198307

PROTEIN DATABANK DEPOSITIONS

 * 1) 1odc STRUCTURE OF ACETYLCHOLINESTERASE (E.C. 3.1.1.7) COMPLEXED WITH N-4'-QUINOLYL-N'-9"-(1",2",3",4"- TETRAHYDROACRIDINYL)-1,8-DIAMINOOCTANE AT 2.2A RESOLUTION
 * 2) 1u65 Ache W. CPT-11
 * 3) 1ut6 STRUCTURE OF ACETYLCHOLINESTERASE (E.C. 3.1.1.7) COMPLEXED WITH N-9-(1',2',3',4'-TETRAHYDROACRIDINYL)-1,8-DIAMINOOCTANE AT 2.4 ANGSTROMS RESOLUTION
 * 4) 1v04 SERUM PARAOXONASE BY DIRECTED EVOLUTION
 * 5) 2ckm TORPEDO CALIFORNICA ACETYLCHOLINESTERASE COMPLEXED WITH ALKYLENE-LINKED BIS-TACRINE DIMER (7 CARBON LINKER)
 * 6) 2cmf TORPEDO CALIFORNICA ACETYLCHOLINESTERASE COMPLEXED WITH ALKYLENE-LINKED BIS-TACRINE DIMER (5 CARBON LINKER)
 * 7) 2j25 PARTIALLY DEGLYCOSYLATED GLUCOCERAMIDASE
 * 8) 2v3d ACID-BETA-GLUCOSIDASE WITH N-BUTYL-DEOXYNOJIRIMYCIN
 * 9) 2v3e ACID-BETA-GLUCOSIDASE WITH N-NONYL-DEOXYNOJIRIMYCIN

MY CURRENT RESEARCH
My PhD research thesis focuses on the enzyme acid-beta-glucosidase, with the emphasis on the enzyme's structure and its interactions with various ligands and their pharmacological potential.

The enzyme is involved in a Gaucher disease, which is the most common lysosomal storage disease, and is associated with mutations in the gene coding for the enzyme acid-beta-glucosidase.

There are ~200 known mis-sense mutations, which result in substitution of amino acids in the protein. Some mutations cause complete deactivation of the enzyme; others impair its stability, and some affect both activity and stability.

There are three known phenotypes of the disease: a mild, severe and acute. The acute phenotype is neuropathic, while severe and mild symptoms are caused by accumulation of glucoceramide in macrophage cells resulting in bone atrophy, spleen enlargement, etc.

We have solved structures of recombinant acid-beta-glucosidase expressed in both mammalian and plant cell cultures, as well as of complexes with molecules, which have been shown to inhibit glucoceramide synthase and in a less extent acid-beta-glucosidase. Yet these molecules were shown to stabilize several mutant forms of the mutant enzyme in Gaucher disease.

We have shown that the molecular basis for the stabilization of acid-beta-glucosidase by these small molecules is by binding to the active site and contributing additional non-covalent bonds that stabilize the enzyme's overall structure.

The research is been performed in laboratories of Prof. Joel L. Sussman, Prof. Israel Silman and Prof. Tony H. Futerman, Weizmann Institute of Science

Last update Boris Brumshtein 00:00, 29 September 2008 (IDT)